초록 ▼

국과수에 도입한 정확도 및 신뢰성이 높은 텐덤질량분석기의 최첨단장비를 이용하여, 혈액중 농약의 정밀미량검색법(microanalysis)을 개발하여 유효화하고 데이터베이스화 하고자하였다. 1차년도에 GC/MS법으로 분석되지 않는 농약을 우선 선정하여 검토하였으며, 이어 혈액에 살서제농약 5종, 우레아계농약 31종을 spiking하여 고상추출법으로 추출하고 LC/MS/MS법으로 분석하여 분석파라미터를 검토하였다. LC/MS/MS 조건으로 이동상, 컬럼, MRM 분석파라미터를 최적화하였다. 검출한계, 직선성, 검출한계, 정량한계 등을 검

국과수에 도입한 정확도 및 신뢰성이 높은 텐덤질량분석기의 최첨단장비를 이용하여, 혈액중 농약의 정밀미량검색법(microanalysis)을 개발하여 유효화하고 데이터베이스화 하고자하였다. 1차년도에 GC/MS법으로 분석되지 않는 농약을 우선 선정하여 검토하였으며, 이어 혈액에 살서제농약 5종, 우레아계농약 31종을 spiking하여 고상추출법으로 추출하고 LC/MS/MS법으로 분석하여 분석파라미터를 검토하였다. LC/MS/MS 조건으로 이동상, 컬럼, MRM 분석파라미터를 최적화하였다. 검출한계, 직선성, 검출한계, 정량한계 등을 검토한 결과 양호한 결과를 도출하였다. 또한 세부연구과제로 이번 연구과제를 통해 사람의 자타살에 많이 이용되는 글리포세이트와 혈중알코올 농도와의 분석 상호작용을 연구하였다.

Abstract ▼

I. The key of research and development
In the case of death from pesticides, not only confirm of pesticides in pesticides bottle and empty bottle at dead scene, but also confirm of pesticides in autopsy sample of a person who has died an unnatural death can be applied investigation cause of death

I. The key of research and development
In the case of death from pesticides, not only confirm of pesticides in pesticides bottle and empty bottle at dead scene, but also confirm of pesticides in autopsy sample of a person who has died an unnatural death can be applied investigation cause of death
II. Background of research and development
All of the autopsy, the percentage of death from pesticides is 1.4% for 6years (2005-2010).
Whether the death from pesticides at unnatural death is carried out by Thin-layer chromatography, color test and GC/MS using stomach contents and pretreated blood, but, by developing microanalysis of pesticides in blood by using TANDEM/MS with high accuracy and reliability, we want to be databased. We want to study for correlation alcohol associated with drinking with glyphosate one of the major pesticide of death of pesticides.
III. Exprimental method
1. In this study, a simple, fast and reliable method for the determination of rodenticides; brodifacoum, bromadiolone, coumatetralyl, difenacoum, flocoumafen and warfarin(I.S.) in postmortem blood was developed and validated. Samples were extracted with ethyl acetate and analyzed by high performance liquid chromatography coupled with electro spray ionization mass spectrometry.
2. To support examination of cause of death in forensic science field, a rapid and accurate method for simultaneous determination of urea pesticide azimsulfuron, bensulfuron, chlorbromuron, chlorotoluron, chlorimuron-ethyl, chlorsulfuron, cinosulfuron, cyclosulfamuron, diflubenzuron, diuron, ethametsulfuron-methyl, ethoxysulfuron, flazasulfuron, flucycloxuron, flufenoxuron, halosulfuron-methyl, iodosulfuron-methyl, imazosulfuron, isoproturon, linuron, methabenzthiazuron, metobromuron, metsulfuron-methyl, monolinuron, nicosulfuron, rimsulfuron, pencycuron, thifensulfuron-methyl, thidiazuron, tribenuron-methyl and triflumuron in blood was developed using liquid chromatography with tandem mass spectrometry. Atrazine-D5 was used as internal standards. The analytical method is based on using mixed mode cartridges through automatic procedure. The pesticides were separated using a on a reversed phase chromatography system consisting of a Synergi 2.5 ㎛ Fusion-RP column(50x2.0 mm, 2.5 ㎛) and then compounds were eluted with a gradient. Pesticides were detected in the positive multiple reaction monitoring mode. The method was validated for the linearity, precision, accuracy for each pesticide. Method validation demonstrated good intra- and inter-assay precision and accuracy for all urea pesticides. The limit of quantification ranged from 10 to 100ng/mL, the linear range of the standard curves was 10 - 1000 ng/mL in blood(r2>0.99).
3. Glyphosate[N-(phosphonomethyl)glycine] is a well-known and extensively used water-soluble herbicide. It is very tricky work to analyze glyphosate in the blood. That’s why there are many analytic methods for the glyphosate by GC, LC, IC, LC-MS-MS etc. We tried simple derivatization method to detect glyphosate and its metabolite AMPA. This new method is rapid and simple, we used double derivatization though. This double derivatization is accomplished with 1st para-toluene sulfonyl chloride(PTSCL) and 2nd TFAA by GC-MS. Generally PTSCL derivatization is an analytic method for glyphosate by HPLC followed by a liquid-liquid extraction(LLE) in acidic conditions. We used the LLE extract further derivatization with TFAA. This double derivatization trial to detect glyphosate was conducted in DW, whole blood and plasma with or without blood alcohol. We can easily derivatize the glyphosate after the liquid liquid extraction. Spectrum for the derivatized glyphosate was same with the direct derivatives of glyphosate with TFA/TFE-OH in the use of SPE extraction. Whole blood was a better matrix for the glyphosate detection than DW. Glyphosate products in the market also have good corelation coefficent (R2) over 0.99. In a real sample, this double derivatization method for glyphosate detection has good result. We are going to apply this method to the another glyphosate cases.
IV. Utilization plan about forensic science investigation
Confirming pesticides at autopsy sample related with homicide and suicide incident about high insurance is essential. Because it is very hard to detect pesticides in blood in case of poisoning with small amount and death in a fall after poisoning, we will take advantage of TANDEM/MS method with lower LOD or LOQ.

목차 Contents

• 연구사업 최종보고서 ... 1
• 성과요약문 ... 2
• Summary ... 3
• 목차 ... 5
• 제1장 연구개발과제의 개요 ... 6
• 제1절 연구개발과제의 목표 ... 6
• 제2절 연구개발과제의 필요성 ... 6
• 제2장 연구개발과제의 국내.외 연구개발 현황 ... 7
• 제3장 연구개발과제의 연구수행 내용 및 결과 ... 7
• [세부과제 1] ... 7
• [세부과제 2] 혈중 글리포세이트 농도에 미치는 알콜의 영향 연구 ... 44
• 제4장 목표달성도 및 관련분야에의 기여도 ... 54
• 제5장 연구개발결과의 활용계획 ... 54
• 제1절 활용성과 ... 54
• 제2절 활용계획 ... 55
• 제6장 참고문헌 ... 55
• 제7장 첨부서류 ... 57
• 끝페이지 ... 58